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. 2009 Dec 21;4(12):e8387. doi: 10.1371/journal.pone.0008387

Figure 1. Purification, aggregation property and activity of FadD13 of M. tuberculosis.

Figure 1

A. Analysis of purified FadD13. The purification was carried out by using Ni-NTA affinity chromatography and analyzed by electrophoresis on a 10% SDS-polyacrylamide gel. M – Molecular weight markers, lane 1- cell free extract, lane 2- unbound proteins, lane 3 – wash with lysis buffer containing 20 mM imidazole, lane 4 – wash with lysis buffer containing 50 mM imidazole, lane 5–8 – elutions with lysis buffer containing 250 mM imidazole. B. Analysis of aggregation property. The aggregating nature of FadD13 was studied by using 7.5% non-reducing non-denaturing polyacrylamide gel. Lane 1–20 µg of the native protein, lane 2–20 µg of the native protein incubated with 10 mM DTT and 1 mM ATP for 2 hours. C. Determination of the enzymatic activity of FadD13. FadD13 assay was carried out by using a radioactivity based TLC assay as described in the “Materials and Methods”. Figure shows the reaction products in the absence (lane 1) and presence (lane 2) of CoenzymeA.